The aims of the proposed research are to isolate RNA-dependent ATPases from the nuclei of animal cells, to characterize the purified enzymes and to determine their functions. The purification will be from calf thymus, calf liver or calf brain, depending on which tissue gives the best yield at the earliest stages of purification. Preliminary evidence indicates that the enzymes are associated with nuclear ribonucleoprotein particles. The distribution of the RNA-dependent activity in the various components of the nucleus will be studied by fractionation of HeLa nuclei containing pulse labeled 3H-RNA to identify the ribonucleoproteins. The function of the ATPase will be determined by testing whether it is a transcription termination factor or a specific ribonuclease that could be involved in RNA processing or whether it is involved in ATP-dependent release of RNA from the nucleus. The transcription and processing studies will be performed using purified adenovirus DNA and wheat germ RNA polymerase II to synthesize primary transcripts. Isolated nuclei from adenovirus-infected cells will be used to study the effects of the RNA-dependent ATPases on release of RNA.